Isolation and purification of a novel antioxidant protein from the water extract of Sundakai ( Solanum torvum) seeds

Abstract

Reactive oxygen species (ROS) at physiological concentrations may be required for normal cell function. Excessive production of ROS can be detrimental to cells, because ROS can cause oxidative damage to lipids, proteins, and DNA. Herein, we describe the isolation and purification of a novel antioxidant protein the water extract of dried, powdered Sundakai ( Solanum torvum [Solanaceae]) seeds. Sundakai belongs to the Solanaceae family, a small shrub, which is distributed widely in India, Malaya, China, Phillipines and tropical America. Fifty percent of ammonium sulphate-precipitated crude water extract was fractionated on a Sephadex G100 column, which yielded two peaks, PI and PII. Peaks PI and PII inhibited lipid peroxidation up to 40% and 89%, respectively in linolenic acid micelles. Rechromatographing of peak PII on Sephadex G100 yielded a single peak, indicating the homogeneity of the purified protein. SDS–PAGE analysis indicated the molecular weight of the purified protein to be ∼28 kDa. The purified protein, at 0.8 μM, inhibited deoxyribose degradation induced by generation of hydroxyl radicals by 90% and scavenged DPPH (1,1-diphenyl-2-picrylhydrazyl) radicals by 76%. The reducing power and chelating power of the purified protein, at 0.8 μM, were found to be 72% and 85%, respectively. The protein, at 0.8 μM, also offered significant protection to calf thymus DNA damage induced by H 2O 2 (1 mM). Therefore, the present study demonstrates, for the first time, a novel protein from the water extract of Sundakai seeds as an excellent antioxidant.